The Program Project's investigational focus is on the multimodal treatment of AJCC Stage III melanoma patients by surgery, active specific immunotherapy (PMCV; polyvalent melanoma cell vaccine), and IFNalpha-2b in a multicenter randomized Phase III clinical trial. The success of this clinical trial requires endpoints for determining treatment efficacy. The duration of disease-free and overall survival are well established measured clinical endpoints but their assessment requires lengthy follow- up. Also, they cannot be used as criteria for monitoring the early response to therapy or predicting recurrent metastatic disease. Project II will focus on investigating prognostic molecular markers (PMM) to assess their clinical utility in identifying disease status and response to therapy before there is any clinically evident change. Prognostic molecular markers will be assessed qualitatively an semi- quantitatively by employing a large-scale multimarker reverse transcriptase-polymerase chain reaction (RT-PCR) and automated Southern blot assay system we have established. RT-PCR and Southern blot assays will be used to: (1) validate PMM for detection of subclinical recurrent disease and as determinants of tumor progression; (2) assess PMM clinical utility in predicting the clinical efficacy of therapeutic interventions in melanoma patients receiving surgery, PMCV and IFNalpha-2b; and, (3) correlate PMM with corresponding specific cell-mediated responses (delayed-type hypersensitivity skin tests) and antibody responses to immunogenic melanoma-associated antigens (MAA). The latter study will be focused on PMM which are known immunogenic MAA and for which recombinant proteins will provide an excellent patient resource to assess the clinical utility of PMM.